In vitro- and ex vivo- investigations of cytokine expression in the equine tracheal epithelium

Titelblatt, Inhaltsverzeichnis, Lebenslauf Einleitung, Literaturübersicht Material und Methoden Ergebnisse Teil 1 Ergebnisse Teil 2 Diskussion Zusammenfassung Summary LiteraturverzeichnisIn dieser Arbeit wurden in vitro- und ex vivo- Untersuchungen zu viralen Infektionen des Trachealepithels des Pferdes und der dadurch ausgelösten Zytokinexpression durchgeführt. Hauptuntersuchungsgegenstand war eine im Institut für Veterinär-Pathologie etablierte Zellkultur aus equinem Trachealepithel (ET Zellen). Die ET Zellen wurden mit den equinen Herpesviren vom Typ 1, 2 und 4 (EHV-1, -2, -4), den equinen Rhinitisviren Perv und 350/72 (equines Rhinitis A Virus: ERAV) und P 1436/73 (equines Rhinitis B Virus: ERBV) sowie mit dem Vesikulären Stomatitisvirus (VSV) und dem Erreger der equinen Arteriitis (EAV) infiziert. Nach Erstinfektion wurden die Zellen zweimal passagiert und es wurden Wachstumskurven für diese Viren erstellt. Außer EHV-4 führten alle verwendeten Viren zu einer produktiven Infektion der ET Zellen. Für ERAV und ERBV ist damit bewiesen, auch die unteren Luftwege des Pferdes zu infizieren und pathogene Mechanismen auslösen zu können. Weiterhin ist die Vermutung über die Bedeutung von EHV-2 für respiratorische Erkrankungen des Pferdes untermauert worden. Interessanterweise kam es für EHV-4, den typischen Erreger der Rhinopneumonitis beim Pferd, nicht zu einer produktiven Infektion der ET Zellen. Möglicherweise handelt es sich um eine restringiert-persistente Infektion, dies muß in weiteren Untersuchungen erforscht werden. Anschließend wurde die Zytokinexpression durch die ET Zellen vor und nach Infektion mit EHV-1, -2, -4 sowie beiden Stämmen der ERAV bestimmt. Dazu wurde nach RNA Isolierung und darauffolgender Umschreibung in cDNA die erhaltene cDNA mit Hilfe zytokinspezifischer Primer vervielfältigt (RT-PCR). Es wurden die Primer für equines Interleukin (IL) -1a, -1b, -2, -4, -5, -6, -8, -10, -11, -13, Tumornekrosefaktor a (TNFa), Interferon g (IFNg) und Granulozyten-Makrophagen-Kolonienstimulierender Faktor (GM-CSF) verwendet. Für die ET Zellen wurde eine Zytokinexpression für IL-1a, -1b, -6, -8, -10 und -11 nachgewiesen. Alle Zytokine wurden schon vor Infektion exprimiert. Nach viraler Infektion konnten Änderungen der Zytokinexpression beobachtet werden. Als ein wichtiges Ergebnis stellte sich heraus, daß beide ERAV deutlich die Zytokinexpression für IL-11 erhöhten. Bei der angewandten RT-PCR handelt es sich um einen qualitativen Nachweis, in weiteren Untersuchungen sollten zur besseren Auswertung auch quantitative Methoden zum Einsatz kommen. Um Aussagen über das Zytokinspektrum in den unteren Atemwegen der Pferde zu erhalten, wurden weiterhin von 15 Schlachtpferden Zellproben durch Abschaben der Trachea und ihrer Bifurkation mit dem Skalpell gewonnen. Dafür wurden Tracheen von Lungen ohne makroskopisch sichtbare pathologische Veränderungen ausgewählt. Es konnte mRNA für IL-8 (100% der Pferde), IL-2 (67%), IL-1b, -6, -10 (60%), IFNg (53%), IL-1a (20%) sowie für IL-11, TNFa und GM-CSF (13%) nachgewiesen werden. mRNA für IL-4, -5 und -13, welche kennzeichnend für Th2-Lymphozyten sind, konnte bei keinem der Pferde festgestellt werden. Das Vorkommen von IL-2 und IFNg bei einem Großteil der Pferde sowie das Fehlen von IL-4, -5 und -13 zeigen, daß bei Pferden ohne morphologische Veränderungen im Epithel der unteren Atemwege eine Th1-Immunlage vorzuherrschen scheint. Die Zellen von vier Pferden wurden nach der Gewinnung sofort mit EHV-1, -2 und -4 sowie mit den equinen Rhinitisviren infiziert. Hier zeigte sich interessanterweise, daß schon die Kultivierung der Zellen allein zu Änderungen in der Zytokinexpression führte. Aussagen über Änderungen der Zytokinexpression nach Infektion konnten ohne Quantifizierung nicht getroffen werden. Mit dieser Arbeit wurde gezeigt, daß dem equinen Trachealepithel aus immunologischer Sicht große Bedeutung für die Pathogenese respiratorischer Erkrankungen zugeschrieben werden kann und es Gegenstand weiterer Forschungsarbeiten werden sollte.These studies were undertaken to explore the role of viral infection of the equine tracheal epithelium and virally induced cytokine expression. The main target was a cell culture obtained from equine tracheal epithelial cells (ET cells). The ET cell line was established in the institute for veterinary pathology. It is the second permanent cell line of equines achieved worldwide. ET cells were infected with equine herpesvirus type 1, 2 and 4 (EHV-1, -2, -4); equine rhinitis virus serotype A (strains Perv and 350/72) and serotype B (strain P 1436/73); vesicular stomatitis virus and with equine arteritis virus. After primary infection, the cells were passaged two times and growth kinetics were evaluated for all viruses. All viruses (except EHV-4) led to productive infection of ET cells. These findings confirm the ability of equine rhinitisviruses to infect the lower airways which should be investigated in following studies. Additionally, the results provide further evidence of the infectious potential of EHV-2 as a respiratory pathogen for the lower airways of the horse. Interestingly EHV-4 was not able to set up a productive infection cycle in ET cells. We presumed a restricted persistent infection which should be subject to further investigation. Virally induced expression of cytokines was determined by using the reverse transcription-polymerse chain reaction (RT-PCR). Total cellular RNA was extracted from ET cells, reverse transcripted in cDNA and amplified with cytokine specific primers for equine interleukin (IL) 1a, 1b, 2, 4, 5, 6, 8, 10, 11, 13, interferon gamma (IFNg), tumour necrosis factor alpha (TNFa) and granulocyte-macrophage colony- stimulating factor (GM-CSF). ET cells expressed mRNA for IL 1a, 1b, 6, 8, 10 and 11. All cytokines could already be measured before infection, but viral infection led to changes in cytokine expression. An important result in this context is, that both equine rhinitis A viruses increased expression for IL 11. In following studies the use of RT-PCR should be combined with quantitative methods. To evaluate cytokine expression in the tracheal epithelium of the horse, tracheal tissue of 15 slaughtered horses without morphological changes in trachea and lungs were sampled. Cells were obtained by gentle scraping off the tracheal epithelium layer with a scalpel. mRNA for equine IL 8 (100% of the horses), IL 2 (67%), IL 1b, 6, 10 (60%), IFNg (53%), IL 1a (20%) and for IL 11, TNFa and GM-CSF (13%) could be detected. mRNA for IL 4, 5 and 13, which are characteristic for an immunreaction of the th2 phenotype was not found in any horse. In contrast, expression of mRNA for IL 2 and IFNg was present in most of the horses. This and the missing IL 4, 5 and 13 detection indicates a th1 phenotype in the tracheal epithelium of horses without morphological changes. Cells of four horses were infected immediately after scraping with EHV-1, -2 and -4 and with equine rhinitis viruses. Interestingly, culturing of cells alone led to changes in cytokine expression. Viral influence of cytokine expression could not be evaluated because of the lack of quantitative measurement. This work shows that the equine tracheal epithelium apparently plays a central role in immunological reactions in the pathogenesis of respirator

In-Depth Interviews With State Public Health Practitioners On The United States National Physical Activity Plan

Abstract Background The United States National Physical Activity Plan (NPAP; 2010), the country’s first national plan for physical activity, provides strategies to increase population-level physical activity to complement the 2008 physical activity guidelines. This study examined state public health practitioner awareness, dissemination, use, challenges, and recommendations for the NPAP. Methods In 2011–2012, we interviewed 27 state practitioners from 25 states. Interviews were recorded and transcribed verbatim. Transcripts were coded using a standard protocol, verified and reconciled by an independent coder, and input into qualitative software to facilitate development of common themes. Results NPAP awareness was high among state practitioners; dissemination to local constituents varied. Development of state-level strategies and goals was the most frequently reported use of the NPAP. Some respondents noted the usefulness of the NPAP for coalitions and local practitioners. Challenges to the plan included implementation cost, complexity, and consistency with other policies. The most frequent recommendation made was to directly link examples of implementation activities to the plan. Conclusions These results provide early evidence of NPAP dissemination and use, along with challenges encountered and suggestions for future iterations. Public health is one of eight sectors in the NPAP. Further efforts are needed to understand uptake and use by other sectors, as well as to monitor long-term relevance, progress, and collaboration across sectors

Multimorbidity patterns and health-related quality of life in Jamaican adults: a cross sectional study exploring potential pathways

IntroductionMultimorbidity and health-related quality of life (HRQoL) are intimately linked. Multiple chronic conditions may adversely affect physical and mental functioning, while poorer HRQoL may contribute to the worsening course of diseases. Understanding mechanisms through which specific combinations of diseases affect HRQoL outcomes can facilitate identification of factors which are amenable to intervention. Jamaica, a middle-income country with high multimorbidity prevalence, has a health service delivery system dominated by public sector provision via a broad healthcare network. This study aims to examine whether multimorbidity classes differentially impact physical and mental dimensions of HRQoL in Jamaicans and quantify indirect effects on the multimorbidity–HRQoL relationship that are mediated by health system factors pertaining to financial healthcare access and service use.Materials and methodsLatent class analysis (LCA) was used to estimate associations between multimorbidity classes and HRQoL outcomes, using latest available data from the nationally representative Jamaica Health and Lifestyle Survey 2007/2008 (N = 2,551). Multimorbidity measurement was based on self-reported presence/absence of 11 non-communicable diseases (NCDs). HRQoL was measured using the 12-item short-form (SF-12) Health Survey. Mediation analyses guided by the counterfactual approach explored indirect effects of insurance coverage and service use on the multimorbidity–HRQoL relationship.ResultsLCA revealed four profiles, including a Relatively Healthy class (52.7%) characterized by little to no morbidity and three multimorbidity classes characterized by specific patterns of NCDs and labelled Metabolic (30.9%), Vascular-Inflammatory (12.2%), and Respiratory (4.2%). Compared to the Relatively Healthy class, Vascular-Inflammatory class membership was associated with lower physical functioning (β = −5.5; p < 0.001); membership in Vascular-Inflammatory (β = −1.7; p < 0.05), and Respiratory (β = −2.5; p < 0.05) classes was associated with lower mental functioning. Significant mediated effects of health service use, on mental functioning, were observed for Vascular-Inflammatory (p < 0.05) and Respiratory (p < 0.05) classes.ConclusionSpecific combinations of diseases differentially impacted HRQoL outcomes in Jamaicans, demonstrating the clinical and epidemiological value of multimorbidity classes for this population, and providing insights that may also be relevant to other settings. To better tailor interventions to support multimorbidity management, additional research is needed to elaborate personal experiences with healthcare and examine how health system factors reinforce or mitigate positive health-seeking behaviours, including timely use of services

Комплексный подход по подбору скважинного оборудования на Ванкорском нефтегазовом месторождении (Красноярский край)

Анализ осложнений при эксплуатации скважин в условиях Ванкорского нефтегазового месторождения. Технологические решения по подбору скважинного оборудования добывающих скважин.Analysis of complications in the operation of wells in the conditions of the Vankor oil and gas field. Technological solutions for the selection of downhole equipment for producing wells

Driven to Support: Individual- and County-Level Factors Associated With Public Support for Active Transportation Policies

Purpose: To assess predictors of stated support for policies promoting physically active transportation. Design: Cross-sectional. Setting: US counties selected on county-level physical activity and obesity health status. Participants: Participants completing random-digit dialed telephone survey (n = 906). Measures: Survey measures assessed stated support for 5 policies to promote physically active transportation, access to active transportation facilities, and time spent in a car. County-level estimates included household car dependence and funding for bicycle–pedestrian projects. Analysis: Multivariable generalized linear mixed models using binary distribution and logit link, accounting for clustering within county. Results: Respondents supported policies for accommodating bicyclists and pedestrians through street improvements (89%), school active transportation programs (75%), employer-funded active commuting incentives (67%), and allocation of public funding (68%) and tax support (56%) for building and maintaining public transit. Residents spending >2 h/d (vs $1.6 million in bicycle and pedestrian improvements expressed greater support for funding (OR: 1.71; CI: 1.04-2.83) and tax increases (OR: 1.73; CI: 1.08-2.75) for transit improvements compared to those with lower prior investments ($1.6 M), public transit is nearby, and respondents drive >2 h/d

Validity and reliability of the semi-quantitative self-report Home Food Availability Inventory Checklist (HFAI-C) in White and South Asian populations.

Despite interest in the importance of the home food environment and its potential influence on children's diets and social norms, there remain few self-report checklist methods that have been validated against the gold standard of researcher-conducted inventories. This study aimed to assess the criterion validity and reliability of the 'Home Food Availability Inventory Checklist' (HFAI-C), a 39-item checklist including categories of fruit, vegetables, snacks and drinks.The HFAI-C was completed by 97 participants of White and Pakistani origin in the UK. Validity was determined by comparing participant-reported HFAI-C responses to data from researcher observations of home food availability using PABAK and weighted kappa statistics. The validity of measuring the amount of items (in addition to presence/absence) available was also determined. Test-retest reliability compared repeated administrations of the HFAI-C using intra-class correlation coefficients.Validity and reliability was fair to moderate overall. For validity, the average category-level PABAK ranged from 0.31 (95 % CI: 0.25, 0.37) for vegetables to 0.44 (95 % CI: 0.40, 0.49) for fruits. Assessment of the presence/absence of items demonstrated higher validity compared to quantity measurements. Reliability was increased when the HFAI-C was repeated close to the time of the first administration. For example, ICCs for reliability of the measurement of fruits were 0.52 (95 %CI: 0.47, 0.56) if re-administered within 5 months, 0.58 (95 % CI: 0.51, 0.64) within 30 days and 0.97 (95 %CI: 0.94, 1.00) if re-administered on the same day.Overall, the HFAI-C demonstrated fair to moderate validity and reliability in a population of White and South Asian participants. This evaluation is consistent with previous work on other checklists in less diverse, more affluent populations. Our research supports the use of the HFAI-C as a useful, albeit imperfect, representation of researcher-conducted inventories. The feasibility of collecting information using the HFAI-C in large, multi-ethnic samples can facilitate examination of home food availability in relation to exposures such as ethnicity and outcomes including behavioural, social and health outcomes. Future work using the HFAI-C could provide important insights into a modifiable influence with potential to impact health

Overweight and obesity in relation to cardiovascular disease risk factors among medical students in Crete, Greece

BACKGROUND: Recent data indicate increasing rates of adult obesity and mortality from cardiovascular disease (CVD) in Greece. No data, however, are available on prevalence of overweight and obesity in relation to CVD risk factors among young adults in Greece. METHODS: A total of 989 third-year medical students (527 men, 462 women), aged 22 ± 2 years, were recruited from the University of Crete during the period 1989–2001. Anthropometric measures and blood chemistries were obtained. The relationships between obesity indices (body mass index [BMI], waist circumference [WC], waist-to-hip ratio [WHpR], waist-to-height ratio [WHtR]) and CVD risk factor variables (blood pressure, glucose, serum lipoproteins) were investigated. RESULTS: Approximately 40% of men and 23% of women had BMI ≥ 25.0 kg/m(2). Central obesity was found in 33.4% (average percentage corresponding to WC ≥ 90 cm, WHpR ≥ 0.9 and WHtR ≥ 50.0) of male and 21.7% (using WC ≥ 80 cm, WHpR ≥ 0.8, WHtR ≥ 50.0) of female students. Subjects above the obesity indices cut-offs had significantly higher values of CVD risk factor variables. BMI was the strongest predictor of hypertension. WHtR in men and WC in women were the most important indicators of dyslipidaemia. CONCLUSION: A substantial proportion of Greek medical students were overweight or obese, obesity status being related to the presence of hypertension and dyslipidaemia. Simple anthropometric indices can be used to identify these CVD risk factors. Our results underscore the need to implement health promotion programmes and perform large-scale epidemiological studies within the general Greek young adult population

Association between television viewing and the risk of metabolic syndrome in a community-based population

<p>Abstract</p> <p>Background</p> <p>As a result of metabolic syndrome becoming an important issue during recent decades, many studies have explored the risk factors contributing to its development. However, less attention has been paid to the risk associated with sedentary behavior, especially television viewing. This study examined the association between television viewing time and the risk of having metabolic syndrome in a population of Taiwanese subjects.</p> <p>Methods</p> <p>This community-based cross-sectional study included 2,353 subjects (1,144 men and 1,209 women) aged 40 and over from October, 2004 to September, 2005. Information about the time spent watching TV was obtained using a self-administered questionnaire. The definition of metabolic syndrome was according to the Third Report of the National Cholesterol Education Program's Adult Treatment Panel modified for Asians.</p> <p>Results</p> <p>Compared to subjects who viewed TV < 14 hr/week, those who viewed TV > 20 hr/week had a 1.50-fold (95% confidence intervals (CI): 1.10, 2.03) risk for men and a 1.93-fold (95% CI: 1.37, 2.71) risk for women of having metabolic syndrome, after adjusting for physical activity and other covariates. Stratifying by the three categories of total activity levels, TV viewing time > 20 hr/week was found to still hold a significant risk for having metabolic syndrome in the lowest of the three categories of total activity level for men and in all three categories of total activity level for women.</p> <p>Conclusion</p> <p>The findings suggest that TV viewing is an independent risk factor associated with metabolic syndrome in Taiwanese people.</p

Type 2 diabetes in adolescents and young adults

The prevalence of type 2 diabetes in adolescents and young adults is dramatically increasing. Similar to older-onset type 2 diabetes, the major predisposing risk factors are obesity, family history, and sedentary lifestyle. Onset of diabetes at a younger age (defined here as up to age 40 years) is associated with longer disease exposure and increased risk for chronic complications. Young-onset type 2 diabetes also affects more individuals of working age, accentuating the adverse societal effects of the disease. Furthermore, evidence is accumulating that young-onset type 2 diabetes has a more aggressive disease phenotype, leading to premature development of complications, with adverse effects on quality of life and unfavourable effects on long-term outcomes, raising the possibility of a future public health catastrophe. In this Review, we describe the epidemiology and existing knowledge regarding pathophysiology, risk factors, complications, and management of type 2 diabetes in adolescents and young adults